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Targeted RNA Sequencing Assay to Characterize Gene Expression and Genomic Alterations
Published on: August 4, 2016
Targeted high-throughput sequencing of tagged nucleic acid samples
Matthias Meyer1, Udo Stenzel, Sean Myles
1Max Planck Institute for Evolutionary Anthropology, Deutscher Platz 6, D-04103 Leipzig, Germany. mmeyer@eva.mpg.de
Nucleic Acids Research
|August 3, 2007
Summary
Parallel tagged sequencing (PTS) enables cost-effective, high-throughput DNA sequencing by barcoding multiple samples. This method significantly boosts the efficiency of 454 sequencing for applications like mitochondrial DNA genome analysis.
Area of Science:
- Genomics and Molecular Biology
- Next-Generation Sequencing Technologies
Background:
- High-throughput 454 DNA sequencing offers speed and cost benefits over Sanger sequencing.
- Current 454 technology has limitations in parallel sample processing capacity.
- Efficient parallelization is crucial for maximizing the utility of advanced sequencing platforms.
Purpose of the Study:
- To introduce a novel barcoding technique for parallel DNA sample sequencing.
- To enhance the throughput of high-throughput sequencing platforms like 454.
- To enable cost-effective and flexible parallel sequencing of diverse nucleic acid samples.
Main Methods:
- Development and implementation of Parallel Tagged Sequencing (PTS), a barcoding strategy.
- Application of PTS to double-stranded nucleic acid samples.
- Demonstration of PTS utility for sequencing contiguous DNA fragments, including mtDNA genomes.
Main Results:
- PTS allows for the parallel sequencing of any number and type of double-stranded nucleic acid samples.
- The technique is highly effective for sequencing contiguous DNA fragments, such as whole mtDNA genomes.
- Potentially, up to 250 mammalian mtDNA genomes can be sequenced in a single 454 GS FLX run using PTS.
Conclusions:
- Parallel Tagged Sequencing (PTS) significantly increases sample sequencing throughput.
- PTS effectively overcomes the parallel processing limitations of 454 sequencing technology.
- This method fully mobilizes the resources of 454 technology for targeted sequencing applications.
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